title: Immobilization of matrix metalloproteinase 8 (MMP-8) for online drug screening.
abstract: Matrix metalloproteinase 8 (MMP-8) has been reported to have a key role in several pathologic conditions, like heart diseases, osteoarthritis, multiple sclerosis, and various other inflammatory conditions. Therefore, there is a great interest regarding the development of MMP-8 selective inhibitors. In the recent years, immobilized enzyme reactors (IMERs) proved to be an efficient alternative to solution-based assays. Besides the recycling of the enzyme, IMER approach allows a simple way to determine affinity data and thus the ranking of inhibiting potency of the compounds under study, especially when coupled to MS. In this study, the immobilization of MMP-8 was investigated in terms of type of support, kinetic parameters, storage and pH stability. Epoxy activated silica resulted the best matrix for the preparation of an immobilized enzyme reactor (IMER) containing human MMP-8. The IMER was successfully used for the online screening of known MMP-8 inhibitors in zonal chromatography and inhibition experiments.
authors.affiliation: Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Pisa, Italy. lcap@ns.dcci.unipi.it
authors.forename: Francesco
authors.initials: F
authors.lastname: Mazzini
authors.forename: Elisa
authors.initials: E
authors.lastname: Nuti
authors.forename: Antonella
authors.initials: A
authors.lastname: Petri
authors.forename: Armando
authors.initials: A
authors.lastname: Rossello
mesh_terms: D019149:Bioreactors; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D020784:Matrix Metalloproteinase 8 / Q000737:chemistry* / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D009930:Organic Chemicals / Q000737:chemistry / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D061965:Matrix Metalloproteinase Inhibitors; D009930:Organic Chemicals; D020784:Matrix Metalloproteinase 8